3′,5-二甲酰和厚朴酚诱导人肾癌NC-65细胞凋亡作用研究

3',5-Diformyl Honokiol Induced Cell Apoptosis in Human Renal Carcinoma NC-65 Cells

目的:通过体外研究厚朴酚衍生物3′,5-二甲酰和厚朴酚(3a)对人肾癌NC-65细胞增殖、凋亡及相关蛋白水平的影响,探讨3a诱导凋亡、抑制肾癌细胞增殖的作用机制。方法:采用CCK-8法检测3a对NC-65细胞生存率的影响;通过Hoechst 33342染色法观察3a刺激细胞24 h后细胞形态学的改变;利用流式细胞术观察3a刺激后细胞凋亡以及周期变化;蛋白免疫印迹技术(Western blotting)检测细胞凋亡及周期相关蛋白的表达水平改变。结果:3a对人肾癌NC-65细胞有明显抑制增殖的作用,呈剂量依赖;3a可使NC-65细胞周期阻滞于G_(0)/G_(1)期;经药物处理后,细胞凋亡率随3a浓度增加呈上升趋势;凋亡相关蛋白Bax、活化的caspase-9、caspase-3蛋白以及抑癌蛋白p53等均表达上调。结论:3a可抑制人肾癌NC-65细胞的增殖,导致细胞周期阻滞,诱导细胞凋亡,其机制可能与调节凋亡及周期相关蛋白的表达有关。

Objective:To investigate the effects of 3',5-diformyl honokiol(3a)on the cell proliferation,apoptosis and related protein expression in human renal carcinoma NC-65 cells and the involved mechanisms.Methods:The survival rate of NC-65 cells was detected by CCK-8 assay;Hoechst 33342 staining was used to observe the morphological changes of cells;The apoptosis and cell cycle were detected with flow cytometry and western blotting was used to detect the expression of the related proteins.Results:The results showed that 3 a could concentration-dependendy inhibit the proliferation of NC-65 cells and induce NC-65 cells to be blocked in G_(0)/G_(1) stage;3a can significantly modulate the expression of protein Bax,cleaved caspase-3 and the expression of other apoptosis-related protein.Conclusion:Honokiol analog 3a could inhibit the proliferation of human renal cell carcinoma NC-65 cells,block the cells in G0/G1 phase and induce cell apoptosis.The mechanism may be that 3a can modulate the expression of apoptosis-related proteins.